Treatment of aspergillus infections with alpha thymosin peptides

ABSTRACT

A method for treating a human infected with  Aspergillus  by using thymosin alpha 1 as an immuno-stimulator in activating dendritic cells. The method is particularly useful in preventing an infection by  Aspergillus  in an immuno-compromised host being treated with a bone marrow transplantation.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of provisional application60/457,911, filed Mar. 28, 2003.

FIELD OF THE INVENTION

The present invention relates to the treatment of fungal infections. Inparticular the present invention relates to the treatment and preventionof Aspergillus infections such as Invasive Aspergillosis associated withbone marrow transplantations.

BACKGROUND OF THE INVENTION

Invasive aspergillosis (IA), characterized by hyphal invasion,destruction of pulmonary tissue and dissemination to other organs, isthe leading cause of both nosocomial pneumonia and death in allogeneicbone marrow transplantation (BMT) with an estimated infection rate of 5to 10% and an associated mortality rate of 90 to 100%. The mostimportant risk factor for IA has historically been neutropenia, suchthat reconstitution with myeloid progenitors offered protection againstIA in a murine model of allogeneic BMT. However, recent studies on theepidemiology of IA in BMT recipients indicated a reducedneutropenia-related infection and an increase “late-onset” infection, inconcomitance with the occurrence of graft versus host disease.

There is a need in the art for methods of treating Asperigillusinfection.

SUMMARY OF THE INVENTION

In accordance with the present invention, a method for treating orpreventing an Aspergillus infection in a mammal comprises administeringto the mammal a pharmaceutical composition comprising an antifungaleffective amount of thymosin alpha 1 (TA1).

DETAILED DESCRIPTION OF THE INVENTION

Clinical and experimental evidence suggest a role of a Th1 cellreactivity in the control of IA. Dendritic cells (DCs) instruct Th1priming to the fungus in vivo and in vitro. Evidence indicates that theability of pulmonary DCs to instruct the appropriate T cell responses tofungal antigens may be affected by local immuno-regulatory events,including signaling through Toll-like receptors (TLRs). DCs may bepromising targets for intervention for immunotherapy and vaccinedevelopment, and shifts the focus of pharmaceutical intervention towardsan “adjuvant”. An adjuvant which is capable of both stimulating theappropriate type of response best tailored to combating the infectionand being effective in conditions of immunosuppression is advantageous.

Thymosin alpha 1 (TA1) is a naturally occurring thymic peptide. In theform of a synthetic 28-amino acid peptide, TA1 is in clinical trialsworldwide for the treatment of some viral infections, either asmonotherapy or in combination with interferon alpha. The treatment ofsome immunodeficiencies, malignancies and HIV/AIDS are additionalindications for TA1. The mechanism of action of a synthetic polypeptideof TA1 is not completely understood but is thought to be related to itsimmuno-modulating activities, centered primarily on the augmentation ofT-cell function. Because of its immunomodulatory function on cells onthe innate immune system, including the ability to activatemitogenactivated protein kinases (MAPKs) and gene expression onmacrophages, we have considered TA1 as an adjuvant capable of activatingDCs for Th1 priming to Aspergillus. The present invention provides atreatment of Aspergillus infections wherein TA1 may activate DCs forantifungal Th1 priming by signaling through TLRs.

The present invention provides a method for treating a mammal infectedwith Aspergillus comprising administering an antifungal effective amountof TA1 to such a mammal. In a preferred embodiment TA1 is effectiveagainst Invasive Aspergillosis (IA). The effective dose of TA1 issufficient to activate dendritic cells to produce Th1 cell promotingcytokines. A preferred dose for treating the fungal infection is in therange between 200 and 400 micrograms/kg body weight per day. In apreferred embodiment the mammal is an immuno-compromised host,particularly a human. The method is particularly useful in treatingimmuno-comprised patients, specifically those patients who are bonemarrow transplantation recipients.

The present invention also provides a method for preventing anAspergillus infection in a mammal comprising administering to suchmammal an antifungal effective amount of TA1. The invention isparticularly useful in preventing IA in an immuno-compromised host. In apreferred embodiment the method prevents such infection inimmuno-compromised patients, specifically those patients being bonemarrow transplantation recipients. The effective dose of TA1 issufficient to activate dendritic cells to produce Th1 cell promotingcytokines. A preferred dose for preventing the fungal infection is inthe range between 200 and 400 micrograms/kg body weight per day.

Without being bound to any particular theory, it is believed that thepresent invention is based on the discovery of a novel immuno-regulatoryactivity of TA1 for the treatment of or protection against anAspergillus infection. TA1 appears to promote the production of theTh1-promoting cytokines IL-12 p70, IL-10, and IFN-alpha, in varioustypes of DCs through a MyD88-dependent pathway.

In TLR-transfected cells, TA1 appears to directly activate TLR9 but notTLR2 signaling, the last being potentiated in response to relevantligands. Therefore, TA1 appears to activate TLR signaling eitherdirectly or indirectly. The data suggest that TA1 may use theTLR2-dependent pathway on myeloid dendritic cells (MDCs) for IL-12 p70production and the TLR9-dependent pathway on plasmacytoid dendriticcells (PDCs) for IFN-alpha and IL-10 production.

As IL-10 production by DCs may be a component of memory protectiveantifungal immunity, balancing the IL-12/IL-10 production on DCs and/ordifferent DC subsets may be a reason for the very essence ofadjuvanticity of TA1 in Aspergillosis.

In a BMT mouse model, TA1 treatment after Aspergillus infection led toan increase in CD4⁺ and CD8⁺ cells, as well as an increase in totalneutrophils. The frequency of Th1 cells (producing IFN-gamma) wereincreased, while the Th2 cells (producing IL-4) were decreased aftertreatment with TM.

Importantly, treatment of BMT mice infected with Aspergillus with TA1led to a dose-responsive reduction in fungal growth in the lungs, and atthe higher doses was able to affect a complete cure of the infection.TA1 was also able to increase the therapeutic efficacy of amphotericinB.

The effects of TA1 on DCs are consistent with its anti-apoptoticactivity. Since DCs are central in the balancing act betweenimmunopathology, immunity and autoimmunity, and PDCs signaling throughTLR9 are present in the thymus, the ability to modulate DC functioningindicates that TA1 is an endogenous regulator of the innate and adaptiveimmune systems acting through TLR utilization. This provides a rationalefor the therapeutic prescription of TA1 in some viral infections, wherePDCs producing IFN-alpha are considered to play a central role. For theproduction of IFN-alpha in these PDCs, TLR9 is essentially required.Moreover, PDCs appear also to participate in immune responses afterhematopoietic cell transplantation, which may explain, among others, thebeneficial effect of TA1 in the immuno-reconstitution in BMT mammals.

TLRs appear to activate the innate immune system not only to assist theadaptive immune system but also for direct antimicrobial effectoractivity. Since TA1 appears to activate DCs for Th1 priming toAspergillus, and also effector neutrophils to an antifungal state, thisfurther indicates the beneficial effect in the treatment of fungalinfections by TA1.

Aspergillus has a unique nature, in that it is a saprophytic funguscolonizing immunocompromised hosts. The present invention providesdeliberate targeting of cells and pathways of cell-mediated immunity andincreases resistance to Aspergillus, wherein TA1 is the adjuvantprogramming the appropriate Th1 reactivity to the fungus throughutilization of the TLR pathway.

The invention is further illustrated by the following example, which isnot to be construed as limiting.

EXAMPLE 1 Animals

Female, 8- to 10-weeks old, BALB/c and C57BL6 mice were from CharlesRiver. NOD/SCID were from The Jackson. Breeding pairs of homozygousTLR2-, TLR9- and MyD88-deficient mice, raised on C57BL6 background, andof homozygous IFN-gamma- and IL-4-deficient mice, raised on BALB/cbackground, were bred under specific-pathogen free conditions.

Microorganism Infections and Treatments

For infection with A. fumigatus, mice were intranasally injected for 3consecutive days with a suspension of 2×10⁷ conidia/20 microlitersaline. For the quantification of fungal growth in the lungs, the chitinassay was used. The chitin content was expressed as micrograms ofglucosamine per organ. The glucosamine content of lungs from uninfectedmice was used as a negative control ranging between 0.80 and 2.25microgram glucosamine/organ. For histological analysis, lungs wereexcised and immediately fixed in formalin. Sections (3 to 4 micron) ofparaffin-embedded tissues were stained with the periodic acid-Schiffprocedure. Thymosin alpha 1 (TA1) and the scrambled polypeptide are aspurified sterile lyophilized acetylated polypeptides with endotoxinlevels less than 0.03 pg/ml, by a standard limulus lysate assay. Thesequences were as follows:Ac-Ser-Asp-Ala-Ala-Val-Asp-Thr-Ser-Ser-Glu-Ile-Thr-Thr-Lys-Asp-Leu-Lys-Glu-Lys-Lys-Glu-Val-Glu-Glu-Ala-Glu-Asn-O(Thymosin alpha 1) andAc-Ala-Lys-Ser-Asp-Val-Lys-Ala-Glu-Thr-Ser-Ser-Glu-Ile-Asp-Thr-Thr-Glu-Leu-Asp-Glu-Lys-Val-Glu-Val-Lys-Ala-Asn-Glu-OH(sThymosin alpha 1). Their lyophilized powders were reconstituted insterile water.

The treatments were as follows; in BMT-mice, TA1, at different dosesadministered intraperitoneally, sthymosin alpha 1, 400 microgram/kgadministered intraperitoneally or human recombinant G-CSF 250microgram/kg administered intravenously, were given daily beginning theday of the BM infusion, in concomitance with the infection andcontinuing for an additional 3 days. Amphotericin B was given daily for3 days in concomitance with the infection, at a dose of 4000microgram/kg, administered intraperitoneally. This dose would cure IA incyclophosphamide-treated mice. Cyclophosphamide, 150 mg/kg administeredintraperitoneally, was given a day before the infection. Incyclophosphamide-treated mice, 400 microgram/kg TA1 was givenintraperitoneally for 5 consecutive days beginning the day of theinfection.

Neutrophil depletion was obtained by treatment with 1 mg ofGr-1-neutralizing RB6-8C5 antibody intravenously, a day before and afterthe infection. The treatment dramatically reduced the number of lungneutrophils but not that of DCs (between 5 to 6×10⁵ CD11c⁺, MHC ClassII⁺, F480-cells before and after treatment). Control mice received anequivalent amount of purified rat IgG2b. FACS analysis of lung cells aday after treatment with cyclophosphamide revealed a profound andlong-lasting (for up to 5 days) leukopenia. The percentages of F480⁺cells (about 20%) and that of CD11c⁺, MHC Class II⁺, F480-DCs (<3%) wereunaffected by treatment.

TLR Ligands

Zymosan was from Saccharomyces cerevisiae, lipoteichoic acid (LTA) fromStaphylococcus aureus, and lipopolysaccharide (LPS) from SalmonellaMinnesota Re 595. The CpG oligonucleotides 1826 and 2006 were provenimmunostimulatory sequences.

Generation of BMT Mice

C57BL6 mice were exposed to a lethal dose of 9 Gy and infused with Tcell-depleted donor cells from BALB/c mice. More than 95% of the micesurvived showing stable, donor type hematopoietic chimerism, as revealedby donor type MHC class I antigen expression on cells from spleens.

Dendritic Cell Isolation and Culture

Blood CD 11c⁺ myeloid DCs (MDCs) were generated from CD 14⁺ mononuclearcells by magnetic cell sorting and cultured for 5 days in Iscove'smodified medium, containing 10% fetal bovine serum, 50 micromolar2-mercaptoethanol, sodium pyruvate (1 mM), 2 mM L-glutamine, HEPES (10mM), and 50 micrograms/ml gentamycin in the presence of 50 ng/ml rHumanGM-CSF and 200 U/ml rHuman IL-4. Immature MDCs were cultured for 24hours with 1000 ng/ml trimeric human CD40 ligand-leucine-zipper fusionprotein to obtain mature DCs. CD 123⁺ plasmacytoid DCs (PDCs) wereisolated using the BDCA-4 isolation kit. Purity of CD 123⁺ cells was>96%.

For mature PDCs, immature DCs were cultured with the trimeric human CD40ligand as above and 10 ng/ml IL-3. FACS analysis revealed that PDCs wereCD123^(bright), CD4⁺, CD45RA⁺ and CD11c⁻ as opposed to MDCscharacterized as being CD1a⁺, CD11c⁺, CD11b⁺, CD4⁺, CD14^(low) and CD8⁻.The expression of HLA Class II, CD80 and CD86 was high in both immatureand mature DCs. Murine lung CD 11c⁺ DCs (between 5 to 7% positive forCD8alpha and between 30 to 35% positive for Gr-1) were isolated bymagnetic cell sorting.

For phagocytosis, DCs were pre-exposed to 100 ng/ml TA1 for 60 minutesand subsequently incubated at 37° C. with Aspergillus conidia for anadditional 60 minutes. The percentage of internalization was calculatedand photographs were taken. In assessing functional maturation andcytokine determination, purified DCs were resuspended in Iscove's medium(with no serum but with polymixin B, to avoid non-specific activation byserum components and endotoxin) and pulsed with 100 ng/ml TA1 for 24hours either alone or together with TLR ligands or unopsonizedAspergillus conidia.

Phenotypic Analysis

Cell surface phenotype was assessed by reacting samples with FITC- orPE-conjugated rat anti-mouse antibodies. Unrelated hisotype matchedantibodies were used as control.

Antifungal Effector Activity

In determining phagocytosis, bronchoalveolar macrophages and peripheralneutrophils were pre-exposed to 100 ng/ml TA1 for 60 minutes andincubated at 37° C. with unopsonized Aspergillus conidia for 60 minutes.In addition, the conidiocidal activity was assessed by determining thenumber of colony forming units and the percentage of colony formingunits inhibition (mean±SE), referred to as conidiocidal activity.

Assay with HEK293 Transfected Cells.

The human embryonic kidney cell line HEK293, wild type or stablytransfected with human TLR2, TLR9 and TLR4/CD1427 were cultured in lowglucose Dulbecco's modified Eagle's medium supplemented with 10% FCS,HEPES (10nM), L-glutamine (2 microgram/ml), and gentamycin (50microgram/ml). Transfectants were additionally supplemented withpuromycin (100 microgram/ml). For stimulation experiments, cells werecultured at a density of 3 to 5×10⁵ cells/wells in 12-well tissueculture plates overnight. Cells were washed and stimulated with 100ng/ml TA1 either alone or together with TLR ligands for 5 h before theassessment of IL-8 production in the supernatants.

Cytokine and Spot Enzyme-Linked Immunosorbent (ELISPOT) Assay

The levels of TNF-alpha, IL-10, IL-12 p70, IFN-alpha and IL-8 in culturesupernatants were determined by Kit ELISAs. The detection limits (pg/ml)of the assays were <3 (human) and <32 (murine) for TNF-alpha, <12(murine) and <5 (human) for IL-10, <16 (murine) and <3 (human) for IL-12p70 and <25 (human) IL-8. For human IFN-alpha <3 ng/ml. For enumerationof cytokine-producing cells, an ELISPOT assay was used on purified CD4⁺T cells and DCs from lungs.

Proliferation Assay by Flow Cytometric Analysis

Proliferation of lung CD4⁺ T lymphocytes stimulated with 10 microgram/mlCon A or heat inactivated conidia in the presence of lung DCs, wasassessed by labeling with CFSE 5(6)-carboxyfluorescein diacetatesuccinimidyl ester.

Reverse Transcriptase (RT)-PCR

Total RNA was extracted from immature DCs pre-treated with 100 ng/ml TA1for 60 minutes followed by the exposure to unopsonized Aspergillusconidia for 60 minutes, as suggested by initial experiments. Synthesisand PCR of cDNA were performed with forward and reverse PCR primers andthe cycles used for murine and human TLRs and HPRT. The synthesized PCRproducts were separated by electrophoresis on 2% agarose gel andvisualized by ethidium bromide staining.

Analysis of p38 and NF-KB Activation

P38 and NF-kB were activated on lung DCs by exposure for 20 minutes at37° C. to Aspergillus conidia and/or 100 ng/ml TA1. Blots of celllysates were incubated with rabbit polyclonal Abs recognizing either theunphosphorylated form of p38 MAPK, or the double-phosphorylated(Thr-180/Tyr-182) p38 MAPK, or Abs specific for the Rel A, 65 kDa DNAbinding subunit of human NF-kB followed by horseradishperoxidase-conjugated goat anti-rabbit IgG, as per manufacturer'sinstructions. Blots were developed with an Enhanced Chemiluminescencedetection kit. Bands were visualized after exposure of the blots to aKodak RX film. To ensure similar protein loading in each lane, thephospho blots were stripped and the membranes were reprobed with Absagainst p38 and NF-kB.

Thymosin Alpha 1 (TA1) Activates Dendritic Cells (DCs)

Previously it has been shown that murine DCs phagocytose Aspergillus invitro and at the site of infection. TA1, but not the scrambled peptideactivates lung DCs for phagocytosis of unopsonized conidia (more thanhyphae), costimulatory antigen expression and cytokine production. Incontrast, Aspergillus conidia alone does not represent a sufficientstimulus to induce the activation of DCs, but the combined exposure toTA1 remarkably increased the expression of MHC Class II antigens, CD86and CD40 molecules and the frequency of IL-12 p70-producing DCs.Interestingly, IL-12 p70-producing DCs are also increased by thymosinalone. TA1 also activates human MDC and PDC subsets. Both immature andmature DC subsets phagocytose conidia. TAI increased the phagocyticactivity of immature DCs, affects the DC morphology (more cytoplasmicprojections can be detected in immature MDCs) and up-regulate the HLAClass II antigens and costimulatory molecule expression in response toconidia. TA1 significantly increase the release of IL-12 p70 in responseto conidia and to zymosan by immature MDCs and that of IL-10 in responseto canidia by immature PDCs. IFN-alpha can be produced by PDCs inresponse to the TLR9 ligand CpG, which production is significantlypotentiated by TA1. In contrast, the scrambled peptide failed toup-regulate Class II antigens and costimulatory molecule expression andto induce cytokine production by DCs in response to conidia.

Together, these data point to a novel, previously undefined,immuno-regulatory role for TA1 in the activation and functioning of DCs.

TA1 Activates the MvD88-Dependent Pathway Through TLR Signaling

TLR signaling occurred in response to Aspergillus conidia, whichmediates functional responses to the fungus. TA1 strongly activates theexpression of TLR2, TLR5, and TLR9 on murine DCs TLR2 and TLR9 are stillactivated upon the combined exposure to conidia and TA1, whereas theexpression of TLR5, whose expression is inhibited. Again, the scrambledpeptide failed to activate TLR2 and TLR9 expression either alone or inresponse to conidia.

The ability of TA1 to activate TLR-dependent signaling is supported bystudies in HEK293 cells transfected with TLR2, TLR9 and TLR4/CD14 bydetermining the IL-8 production in response to TA1 alone or togetherwith the relevant TLR ligands. In such HEK293 cells TA1 significantlyincreased the production of IL-8 by TLR9-transfected cells either aloneor in response to the TLR9 ligand CpG. However, TA1 did not stimulatethe production of IL-8 by TLR2-transfected cells alone but slightlyincreased the production of IL-8 in these cells in response to zymosan.Furthermore, TA1 did not induce IL-8 in TLR4/CD14-transfected cellseither alone or in response to the TLR4 ligand LPS. TA1 also affects theability of murine DCs to produce IL-12 p70 and IL-10 in response tothese microbial TLR ligands. TA1 did not affect cytokine production inresponse to Poly(I:C) or LPS (TLR4), TA1 significantly increased theproduction of IL-12 p70 and decreases that of IL-10 after stimulationwith zymosan and LTA (TLR2) and CpG (TLR9). Therefore, TA1 appears to beable to signal directly through TLR9 and to potentiate TLR2 signaling bythe relevant ligand.

Both NF- and p38 MAPK activation are early events in triggeringTLR-induced gene expression, and TA1 has been previously shown toactivate MAPK-transduction pathways. In support of its involvement inthe TLR-induced pathways, TA1 induced the nuclear translocation of NF-kBas well as p38 phosphorylation (which were not stimulated by eitherconidia alone, the scrambled peptide, or the scrambled peptide plusconidia). Furthermore, inhibitors of NF-kB nuclear translocation (SN50)or p38 MAPK (SB202190 ablate the effect of TA1 on DCs.

The myeloid differentiation factor 88 (MyD88) is one of the adaptorprotein essential for the activation of NF-kB and MAPK and theproduction of IL-12 p70 upon signaling by TLRs. The effect of TA1 andconidia on IL-12 p70 production, and the effect of TAI1 on IL-10production are dramatically ablated in MyD88-deficient mice. Therefore,the MyD88-dependent pathway appears to play an essential role in themechanism of action of TA1 in vitro. To determine whether theMyD88-dependent pathway plays an essential role in TA1 action in vivo aswell. Local fungal growth was assessed after infection of wild type.TLR2-, TRL9- or MyD88-deficient mice with Aspergillus. Fungal growth inTLR2- and TLR9-deficient mice was comparable to that of wild type miceand it is similarly impaired upon thymosin treatment. Fungal growth iscomparable MyD88-deficient mice as well, but in these mice it was notimpaired upon treatment with TA1. Thus, despite a degree of redundancyin the TLR usage, the MyD88-dependent signaling pathway appears to beessential for in the activity of TA1 both in vitro and in vivo.

TA1 Protects BMT-Mice from IA

Treatment with TA1, but not with the scrambled peptide, appeared to beable to cure BMT mice with IA, as revealed by increased survival thatparallels reduced fungal growth in the lungs. The effect on protectionis dose-dependent full protection (>60 d survival) being achieved inmice treated with 200 and 400 microgram/kg TA1 and is superior to thatof amphotericin B. Moreover, TA1 increases the therapeutic efficacy ofamphotericin B, as indicated by the increased survival and decreasedfungal burden of mice treated with both agents. Furthermore, TA1 alsodecreases lung pathology. Lung sections from infected mice show thepresence of numerous Aspergillus hyphae infiltrating the lungparenchyma, with severe signs of bronchial wall damage and necrosis andscarce inflammatory cell recruitment. In contrast, these features arenot observed in TA1 treated mice, whose lungs are characterized byhealing infiltrates of inflammatory cells with no evidence of fungalgrowth and bronchial wall destruction. Thus, TA1 may have therapeuticefficacy in IA and may be beneficial in combination with antifungalsknown to have a reduced activity in BMT settings.

TA1 Accelerates Myeloid and Th1 Cell Recovery in Mice with IA

The absolute number of circulating lymphocytes and neutrophilssignificantly increases after TA1 treatment. More importantly, as bloodneutrophil levels do not predict susceptibility to aspergillosis. Incytofluorimetric analysis however the numbers of lung CD4⁺ and CD8⁺cells and neutrophils were significantly increased upon treatment of BMTmice with TA1. These lung CD4⁺ T lymphocytes are functionally active asindicated by antigenspecific proliferation and IFN-gamma production. Thefrequency of Th1 cells (producing IFN-gamma) producing cells is higher,and Th2 cells (producing IL-4) is lower in mice treated with TA1.Further, the with respect to antifungal activity of effector phagocytes,the conidiocidal activity of both macrophages and neutrophils is higherin TA1 treated mice. Therefore, TA1 appears to not only promote DCmaturation but also to activate local effector cells for promptphagocytosis and killing of the fungus.

Recovery from neutropenia alone, for example by treatment with a dose ofG-CSF known to accelerate neutrophil recovery in mice, is not sufficientto mediate a degree of antifungal resistance comparable to that obtainedwith TA1. Similarly, despite a significant neutrophil recovery, thetherapeutic efficacy of TA1 in mice devoid of T cells orIFN-gamma-producing Th1 cells is not as great. Furthermore, improvedtherapeutic efficacy of TA1 is achieved in the presence of increased Th1cells, such as that occurring in IL-4-deficient mice. Therefore,although neutrophils play an essential role in medicating antifungalresistance in the absence of an adaptive Th1-dependent immunity, theachievement of a state of full protection to the fungus, as appears tobe obtained by treatment with TA1, may rely on the coordinated actionbetween innate effector phagocytes and protective Th1 cells.

1. A method for treating or preventing an Aspergillus infection in amammal comprising administering to said mammal a pharmaceuticalcomposition comprising antifungal effective amount of thymosin alpha 1TA1).
 2. The method according to claim 1, wherein said TA1 isadministered at a dose sufficient to activate dendritic cells to produceTh1 cell promoting cytokines.
 3. The method according to claim 1,wherein said TA1 is administered at a dose of 200 to 400 micrograms/kgbody weight/day.
 4. The method according to claim 1, wherein said mammalis an immuno-compromised host.
 5. The method according to claim 4,wherein said mammal is a human.
 6. The method according to claim 5,wherein said human is a bone marrow transplantation recipient.
 7. Themethod according to claim 5, wherein said TA1 is administered toactivate dendritic cells to produce Th1 cell promoting cytokines.
 8. Themethod according to claim 5, wherein said TA1 is administered at a doseof 200 to 400 micrograms/kg body weight/day.
 9. The method according toclaim 1, wherein the method further comprises administering to saidperson at least one additional antifungal agent.
 10. The methodaccording to claim 9, wherein the additional antifungal agent isAmphotericin B.
 11. The method according to claim 10, wherein saidAmphotericin B is administered at a dose of 4000 micrograms/kg bodyweight/day.
 12. The method of claim 1 wherein said Aspergillus infectionis Invasive Aspergillosis.